Hi all,
I am not sure if I should ask it here, still new to NGS, and I have lots of questions that I couldn't figured out.
1) I am using a custom adapter (that contains p5 p7 for flow cell binding), custom sequencing primer and also a custom index i7 primer for my work. I come across this dual-indexing, I am just wondering if it is possible to have a custom index i5 primer?
2) I am not familiar with Nextera, why is the dark cycles necessary? why not just custom make an index i5 primer that spam the 7bp constant read and read directly into the index? or wait did I miss out something. where is this index i5 primer primed from?
3) must it be index read 1 = i7 index; index read 2 = i5 index? can't be other way round? it's still on the same strand right?
sorry if I asked too many of the dumb questions.
cheers.
I am not sure if I should ask it here, still new to NGS, and I have lots of questions that I couldn't figured out.
1) I am using a custom adapter (that contains p5 p7 for flow cell binding), custom sequencing primer and also a custom index i7 primer for my work. I come across this dual-indexing, I am just wondering if it is possible to have a custom index i5 primer?
2) I am not familiar with Nextera, why is the dark cycles necessary? why not just custom make an index i5 primer that spam the 7bp constant read and read directly into the index? or wait did I miss out something. where is this index i5 primer primed from?
3) must it be index read 1 = i7 index; index read 2 = i5 index? can't be other way round? it's still on the same strand right?
sorry if I asked too many of the dumb questions.
cheers.
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